WO2016112268A1 - Particules radioluminescentes pour l'amélioration de la radiothérapie du cancer - Google Patents

Particules radioluminescentes pour l'amélioration de la radiothérapie du cancer Download PDF

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WO2016112268A1
WO2016112268A1 PCT/US2016/012616 US2016012616W WO2016112268A1 WO 2016112268 A1 WO2016112268 A1 WO 2016112268A1 US 2016012616 W US2016012616 W US 2016012616W WO 2016112268 A1 WO2016112268 A1 WO 2016112268A1
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cancer
radiation
particle
tumor
radio
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Inventor
You-Yeon Won
Jaewon Lee
Sung Duk JO
Min Kyung JOO
Beom Suk LEE
Sang Yoon KIM
Sun Hwa Kim
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Purdue Research Foundation
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Purdue Research Foundation
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/12—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by a special physical form, e.g. emulsion, microcapsules, liposomes, characterized by a special physical form, e.g. emulsions, dispersions, microcapsules
    • A61K51/1241—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by a special physical form, e.g. emulsion, microcapsules, liposomes, characterized by a special physical form, e.g. emulsions, dispersions, microcapsules particles, powders, lyophilizates, adsorbates, e.g. polymers or resins for adsorption or ion-exchange resins
    • A61K51/1244—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by a special physical form, e.g. emulsion, microcapsules, liposomes, characterized by a special physical form, e.g. emulsions, dispersions, microcapsules particles, powders, lyophilizates, adsorbates, e.g. polymers or resins for adsorption or ion-exchange resins microparticles or nanoparticles, e.g. polymeric nanoparticles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0038—Radiosensitizing, i.e. administration of pharmaceutical agents that enhance the effect of radiotherapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/10—X-ray therapy; Gamma-ray therapy; Particle-irradiation therapy
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00—Luminescent materials, e.g. electroluminescent or chemiluminescent
    • C09K11/02—Use of particular materials as binders, particle coatings or suspension media therefor
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00—Luminescent materials, e.g. electroluminescent or chemiluminescent
    • C09K11/08—Luminescent materials, e.g. electroluminescent or chemiluminescent containing inorganic luminescent materials
    • C09K11/67—Luminescent materials, e.g. electroluminescent or chemiluminescent containing inorganic luminescent materials containing refractory metals
    • C09K11/68—Luminescent materials, e.g. electroluminescent or chemiluminescent containing inorganic luminescent materials containing refractory metals containing chromium, molybdenum or tungsten
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N2005/0658—Radiation therapy using light characterised by the wavelength of light used
    • A61N2005/0661—Radiation therapy using light characterised by the wavelength of light used ultraviolet
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/10—X-ray therapy; Gamma-ray therapy; Particle-irradiation therapy
    • A61N2005/1092—Details
    • A61N2005/1098—Enhancing the effect of the particle by an injected agent or implanted device

Definitions

  • the invention relates generally to a radio-sensitization method to enhance the effectiveness of radiation therapy for treatment of cancer. Specifically, the invention relates to the use of radio-luminescent particles for sensitizing tumor cells to radiation therapy.
  • Radio- sensitizers chemical agents that make cancer cells more sensitive to radiation therapy.
  • UV light in killing cells have been known for more than 50 years.
  • UV has never before been thought useful for radio-sensitization in clinical radiation therapy because UV light has a very limited penetration distance in tissue ( ⁇ 1 mm in human tissue).
  • a method of treating cancer in a subject includes providing radio-luminescent particles to a tumor site associated with said cancer in said subject; and exposing said tumor cells to ionizing radiation, wherein said particles produce ultraviolet (UV) light in response to said ionizing radiation, thereby treating said cancer in said subject.
  • the treatment improves tumor cell killing without increasing toxicity to normal tissue.
  • the particle is a calcium tungstate (CaWC ) particle.
  • the calcium tungstate particle provided to said tumor in said subject at a concentration less than about 1.0 mg/ml.
  • the mean diameter of said particle material is in the range between about 0.1 and 1000 nm.
  • the mean diameter of said particle is ranged between about 1 and 10 ⁇ . In yet another aspect, the mean diameter of said particle is in the range between about 1 and 200 nm.
  • the radiation is high-energy photon radiation. In yet another aspect, the radiation is gamma ( ⁇ ) ray radiation. In yet another aspect, the radiation is X-ray radiation.
  • the particle is functionalized or conjugated with a target agent specific to said cancer. In yet another aspect, the target agent is a biological molecule having specific affinity to said cancer cell so as to enhance the delivery of said particles to said tumor cells. In yet another aspect, the particle is functionalized with folic acid.
  • the particle is functionalized with polyethylene glycol (PEG), poly(D,L-lactic acid-ran-glycolic acid) (PLGA), or a combination thereof.
  • the cancer is lung cancer.
  • the cancer is small-cell lung carcinoma (SCLC).
  • SCLC small-cell lung carcinoma
  • NSCLC non-small-cell lung carcinoma
  • a method of improving the effectiveness of radiation therapy for treatment of cancer in a subject includes providing radio-luminescent particles to a tumor site associated with said cancer in said subject; and exposing said tumor cells to ionizing radiation, wherein said particles emit ultraviolet (UV) light in response to said ionizing radiation, thereby improving the effectiveness of said radiation therapy for treatment of said cancer in said subject.
  • UV ultraviolet
  • a method of sensitizing tumor cells to radiation therapy for treatment of cancer in a subject which includes providing radio-luminescent particles to said tumor; and exposing said tumor to ionizing radiation, wherein said particles emit ultraviolet (UV) light in response to said ionizing radiation, thereby sensitizing said tumor cells to said radiation therapy for treatment of said cancer in said subject.
  • UV ultraviolet
  • a method of improving tumor cell killing for treatment of cancer in a subject which includes providing radio-luminescent particles to a tumor site associated with said cancer in said subject; and exposing said tumor cells to ionizing radiation, wherein said particles emit ultraviolet (UV) light in response to said ionizing radiation, thereby improving tumor cell killing for treatment of said cancer in said subject.
  • UV ultraviolet
  • a method of generating secondary UV radiation in deep tissue tumors which includes delivering radio-luminescent particles to said tumor and illuminating said particles with deep-penetrating radiation, thereby generating said secondary UV radiation in said tumor.
  • a method of treating cancer in a subject which includes providing radio-luminescent particles or particle aggregates to a tumor site associated with said cancer in said subject; and exposing said tumor cells to ionizing radiation, wherein said particles or particle aggregates produce ultraviolet (UV) light in response to said ionizing radiation, thereby enhancing treatment of said cancer in said subject.
  • the treatment improves tumor cell killing without increasing toxicity to normal tissue.
  • the particle comprises a radio-luminescent metal tungstate crystallite material in the form of Mx(W0 4 )y where the metal component (M) can be any compound selected from the "Alkaline Earth Metal", “Transition Metal” or “Poor Metal” group of elements in the periodic table, or an atomic mixture thereof.
  • the particle is a composite material comprising a radio-luminescent metal tungstate crystallite in claim 24 and other biocompatible organic or inorganic compound.
  • the particle comprises a radio-luminescent metal molybdate crystallite material in the form of M x (Mo0 4 ) y where the metal component (M) can be any compound selected from the "Alkaline Earth Metal", “Transition Metal” or “Poor Metal” group of elements in the periodic table, or an atomic mixture thereof.
  • the particle is a composite material comprising a radio- luminescent metal tungstate crystallite in claim 26 and other biocompatible organic or inorganic compound.
  • the particle comprises a radio-luminescent calcium tungstate (CaW0 4 ) crystallite.
  • the particle is a composite material comprising a radio-luminescent calcium tungstate crystallite and other biocompatible organic or inorganic compound.
  • the mean largest dimension of said particle material is in the range between about 1 and 50,000 nm in its unaggregated state. In yet another aspect, the mean largest dimension of said particle aggregate is ranged between about 10 and 500,000 nm. In yet another aspect, the mean diameter of said particle or particle aggregate is in the range between about 1 and 500 nm.
  • the ionizing radiation is high-energy photon radiation.
  • the ionizing radiation is gamma ( ⁇ ) ray radiation.
  • the ionizing radiation is X-ray radiation.
  • the ionizing radiation is electron beam radiation.
  • the ionizing radiation is alpha particles emitted in radioactive decay.
  • the ionizing radiation is beta particles emitted in radioactive decay.
  • the ionizing radiation has a peak energy between about 1 and 50,000 keV.
  • the particle or particle aggregate is functionalized or conjugated through an organic or inorganic linker with a target agent specific to said cancer.
  • the target agent is a biological molecule having specific affinity to said cancer cell so as to enhance the delivery of said particles or particle aggregates to said tumor cells and also the internalization of said particles or particle aggregates by said tumor cells.
  • the particle or particle aggregate is functionalized or coupled through an organic or inorganic linker with folic acid.
  • the particle or particle aggregate is encapsulated within a biocompatible coating material. In yet another aspect, the particle or particle aggregate is encapsulated with a biocompatible amphiphilic block copolymer. In yet another aspect, the particle is coated with polyethylene glycol (PEG), poly(D,L-lactic acid) (PLA), or a copolymeric combination thereof. In yet another aspect, the particle is coated with polyethylene glycol (PEG), poly(D,L-lactic acid-ran-glycolic acid) (PLGA), or a copolymeric combination thereof. In yet another aspect, the particle is coated with polyethylene glycol (PEG), poly(s-caprolactone) (PCL), or a copolymeric combination thereof.
  • the particle is coated with polyethylene glycol (PEG), poly(styrene) (PS), or a copolymeric combination thereof.
  • PEG polyethylene glycol
  • PS poly(styrene)
  • the cancer is a solid tumor.
  • the cancer is a hematological tumor.
  • the cancer is head and neck, breast, prostate, lung, gynecological, cervical or brain cancer.
  • the subject is a human patient. In yet another aspect, the subject is an animal patient. In yet another aspect, the radio-luminescent particles or particle aggregates are administered to the tumor site associated with said cancer in said subject through local, intratumoral, intravenous, intraarterial or intraperitoneal routes.
  • a method of improving the effectiveness of radiation therapy for treatment of cancer in a subject includes providing radio-luminescent particles or particle aggregates to a tumor site associated with said cancer in said subject; and exposing said tumor cells to ionizing radiation, wherein said particles emit ultraviolet (UV) light in response to said ionizing radiation, thereby improving the effectiveness of said radiation therapy for treatment of said cancer in said subject.
  • UV ultraviolet
  • a method of sensitizing tumor cells to radiation therapy for treatment of cancer in a subject which includes providing radio-luminescent particles or particle aggregates to said tumor; and exposing said tumor to ionizing radiation, wherein said particles emit ultraviolet (UV) light in response to said ionizing radiation, thereby sensitizing said tumor cells to said radiation therapy for treatment of said cancer in said subject.
  • UV ultraviolet
  • a method of improving tumor cell killing for treatment of cancer in a subject which includes providing radio-luminescent particles or particle aggregates to a tumor site associated with said cancer in said subject; and exposing said tumor cells to ionizing radiation, wherein said particles emit ultraviolet (UV) light in response to said ionizing radiation, thereby improving tumor cell killing for treatment of said cancer in said subject.
  • UV ultraviolet
  • a method of generating secondary UV radiation in deep tissue tumors which includes delivering radio-luminescent particles or particle aggregates to said tumor and illuminating said particles with deep-penetrating radiation, thereby generating said secondary UV radiation in said tumor.
  • FIG. 1 Schematic illustration of the concept of combining ⁇ radiation therapy with UV treatment through the use of radio-luminescent agents.
  • UV light can be generated in deep tissue tumors by delivering radio-luminescent particles (RLP) to the tumor and illuminating them with deep-penetrating ⁇ rays.
  • RLP radio-luminescent particles
  • UV-induced G2/M arrest makes cancer cells more susceptible to ⁇ radiation damage.
  • FIG. 1 SCC7 cells (grown to about 70% confluency in RPMI 1640 medium (Invitrogen) containing 10% FBS and 100 mg/ml penicilin/streptomycin) were exposed to various radiation environments, ⁇ : 6-MV ⁇ radiation (Clinac 600C, Varian) for 150 seconds (5 Gy total dose).
  • RNP in the presence of added 0.5 mg/ml CaW0 4 radio-luminescent powder (RLP) (> 10 ⁇ diameter) with no ⁇ radiation, ⁇ + RLP: 6-MV ⁇ radiation at 5 Gy total dose in the presence of 0.5 mg/ml CaW0 4 RLP.
  • UV 365 nm wavelength UV irradiation (20 mW/cm 2 power density) for 20 minutes.
  • UV ⁇ ⁇ 20-minute UV irradiation followed by 5-Gy ⁇ radiation
  • ⁇ UV 5-Gy ⁇ radiation followed by 20-minute UV irradiation.
  • Control no ⁇ ray, UV light or RLP treatment.
  • the apoptotic, early apoptotic and necrotic populations were measured as the percentages of total cell populations by FACS with Annexin V/PI double staining. The original two- dimensional dot plots demonstrating the fluorescence gating criteria used are presented in Figure 4.
  • FIG. Photographs of stirred suspensions of CaW0 4 RLP at various concentrations in PBS.
  • FIG. 4 FACS analysis with Annexin V/PI staining.
  • Ql necrosis.
  • Q2 apoptosis.
  • Q3 live.
  • Q4 early apoptosis. See the figure caption of Figure 2 for further details.
  • Figures 5A - 5G Schematic illustration of a PEGylated (i.e., polyethylene glycol-block-D,L-lactic acid) or PEG-PLA-encapsulated) CaW0 4 (CWO) microparticle/nanoparticle.
  • Figure 5B A representative TEM micrograph, ( Figure 5C) the X-ray diffraction pattern, and ( Figure 5D) luminescence emission spectra (under 200 nm excitation) of PLA-PEG-encapsulated CWO microparticles (MPs) and nanoparticles (NPs). All measurements were performed at an identical CWO concentration of 0.1 mg/ml.
  • FIG. 5E A representative dynamic light scattering (DLS) correlation function for PEG-PLA-coated CWO NPs. From this data the mean hydrodynamic diameter of the PEG-PLA-coated CWO NPs was estimated to be about 180 nm. The average diameter of the pristine CWO NPs was about 10 nm (as shown in ( Figure 5B)). The number- average degrees of polymerization of the PEG and PLA blocks of the PEG-PLA block copolymer used were determined by 3 ⁇ 4 NMR spectroscopy to be 113 and 44, respectively.
  • DLS dynamic light scattering
  • FIG. 5F A photograph of PEG-PLA-coated CWO NPs (1.0 mg/ml in Milli-Q water) under 6 MV X-ray irradiation demonstrating the radio- luminescence of CWO.
  • Figure 5G Luminscence emission spectra of uncoated CWO MP suspensions recorded with 200 nm excitation. At high particle concentrations (> 1 mg/ml), optical turbidity does not seem to compromise the luminescence signal.
  • the mean hydrodynamic diameter of the PEG-PLA-coated CWO NPs was about 180 nm ( Figure 6F).
  • the number- average degrees of polymerization of the PEG and PLA blocks of the PEG-PLA block copolymer used were determined by 3 ⁇ 4 NMR spectroscopy to be 113 and 44, respectively.
  • HN31 cells were seeded on 96- well culture plates at a density of 0.5 x 10 4 per well and incubated for 24 h prior to exposure to CWO. The cells were treated with uncoated CWO MPs or PEGylated CWO NPs for 24 h at the various CWO concentrations indicated above. CCK-8 assays were performed at 24 h post-CWO treatment.
  • Figure 7 Effect of ⁇ ray dose on the viability of p53-mutant human head and neck cancer HN31 cells (seeded on 60-mm 2 culture plates at densities of 4 x 10 5 , 2 x 10 5 and 1 x 10 5 cells per plate (for 24, 48 and 72-hour experiments, respectively) with DMEM medium containing 10% FBS, 100 U/ml penicillin and 100 ⁇ g/ml streptomycin). After 24 h incubation the cells were exposed to various doses of 137 Cs ⁇ radiation (IBL 437C, CSI Bio International, France) at a rate of 1 Gy per every 11 s.
  • IBL 437C CSI Bio International, France
  • the apoptotic, early apoptotic and necrotic populations were measured as the percentages of total cell populations by FACS (fluorescence-activated cell sorting) with Annexin V/PI double staining.
  • FIG. 8 Effect of ⁇ irradiation on clonogenic survival of HN31 cells at various radiation doses, ⁇ : ⁇ radiation.
  • MP + ⁇ ⁇ radiation in the presence of added 0.5 mg/ml uncoated CaWC (CWO) radio-luminescent microparticles (MPs) (2 - 3 ⁇ diameter).
  • MPs radio-luminescent microparticles
  • NP + ⁇ ⁇ radiation in the presence of added 0.125 mg/ml PEG-PLA-encapsulated CWO nanoparticles (NPs) (180 nm hydrodynamic diameter).
  • UV ⁇ ⁇ 365 nm wavelength UV irradiation (20 mW/cm 2 power density) for 20 minutes followed by ⁇ radiation, ⁇ UV: ⁇ radiation followed by 20-minute UV irradiation.
  • the values of the Sensitization Enhancement Ratio or SER (defined as the ratio of the radiation dose at 10% clonogenic survival in the absence of CWO relative to the radiation dose at 10% survival in the presence CWO) were estimated to be 1.15 for MP + ⁇ (0.5 mg/ml CWO) and 1.13 for NP + ⁇ (0.125 mg/ml CWO).
  • the p-values relative to control (“ ⁇ ") were estimated to be 0.13 at 3 Gy, 0.04 at 6 Gy, and 0.04 at 9 Gy.
  • the p-values relative to ⁇ were 0.03 at 3 Gy, 0.11 at 6 Gy, and 0.06 at 9 Gy.
  • FIG. 9 Assessment of the radio-sensitization efficacy of intratumorally injected CWO MPs and NPs in mouse HN31 xenografts.
  • Figure 10 Apoptotic and necrotic populations measured by FACS with Annexin V/PI double staining in HN31 cells following exposure to ⁇ radiation (5 Gy) in the absence and presence of concomitant UV light generated by CaW0 4 (CWO). Control: no ⁇ ray, UV light or CWO treatment.
  • MP in the presence of added 0.125 mg/ml uncoated CWO radio- luminescent microparticles (MPs) (2 - 3 ⁇ diameter) with no ⁇ radiation.
  • MPs in the presence of added 0.125 mg/ml PEG-PLA-encapsulated CWO radio-luminescent nanoparticles (NPs) (180 nm hydrodynamic diameter) with no ⁇ radiation, ⁇ : ⁇ radiation.
  • HN31 cells were seeded on 60-mm 2 culture plates at densities of 4 x 10 5 , 2 x 10 5 and 1 x 10 5 cells per plate (for 24, 48 and 72-hour experiments, respectively) with DMEM medium containing 10% FBS, 100 U/ml penicillin and 100 ⁇ g/ml streptomycin). After 24 h incubation the cells were exposed to a total 5-Gy dose of 137 Cs ⁇ radiation (IBL 437C, CSI Bio International, France) at a rate of 1 Gy per every 11 s.
  • IBL 437C CSI Bio International, France
  • FIG. 11 FACS analysis of HN31 cells with Annexin V/PI staining.
  • Ql necrosis.
  • Q2 apoptosis.
  • Q3 live.
  • Q4 early apoptosis. See the figure description of Figure 8 for further details.
  • FIG. 12 Visualization of senescent HN31 cells by ⁇ -galactosidase assay following exposure to ⁇ radiation (5 Gy) in the absence and presence of concomitant UV light generated by CaW0 4 (CWO).
  • Control no ⁇ ray, UV light or CWO treatment
  • ⁇ ⁇ radiation.
  • MP in the presence of added 0.125 mg/ml uncoated CWO radio-luminescent microparticles (MPs) (2 - 3 ⁇ diameter) with no ⁇ radiation.
  • MP + ⁇ ⁇ radiation in the presence of added 0.125 mg/ml uncoated CWO MPs.
  • UV 365 nm wavelength UV irradiation (20 mW/cm 2 power density) for 20 minutes with no ⁇ radiation.
  • UV ⁇ ⁇ 365 nm wavelength UV irradiation for 20 minutes followed by ⁇ radiation
  • ⁇ UV ⁇ radiation followed by 20-minute UV irradiation.
  • HN31 cells total 4 x 10 5 cells
  • IBL 437C 137 Cs ⁇ radiation
  • CSI Bio International France
  • Irradiated cells were cultured for 3 or 6 days. Afterward the cells were stained with X-Gal Staining Solution (Sigma- Aldrich). The cells were imaged using an microscope in order to count blue-stained and unstained cells.
  • the invention relates to a radio-sensitization method to enhance the effectiveness of radiation therapy for treatment of cancer. Specifically, the invention relates to the use of radio- luminescent particles for sensitizing tumor cells to radiation therapy.
  • the invention demonstrates that secondary UV radiation can be generated in deep tissue tumors by delivering radio-luminescent particles to the tumors and illuminating them with deep-penetrating ⁇ rays or X-rays.
  • radio-luminescent particle capable of emitting UV light in response to high-energy ionizing radiation
  • radio-luminescent materials are also referred to as "scintillation crystals”.
  • the radio-luminescent material is calcium tungstate (CaW0 4 ) powder (i.e., particles having diameters greater than a micrometer).
  • the radio- luminescent material is submicrometer- sized CaW0 4 particles (i.e., nanoparticles).
  • radio-luminescent materials that can be used include other types of metal tungstates, i.e., M x (W0 4 ) y where the metal component (M) can be any compound selected from the "Alkaline Earth Metal", “Transition Metal” or “Poor Metal” group of elements in the periodic table, or an atomic mixture thereof.
  • Alkaline Earth Metals refers to any one of or a combination of the elements found in Group 2 of the periodic table as defined in accordance with the characterization set forth by the Royal Society of Chemistry (e.g., Be, Mg, Ca, Sr, Ba).
  • Transition Metal refers to a metal as defined by the IUPAC Gold Book as "an element whose atom has an incomplete d sub-shell, or which can give rise to cations with an incomplete d sub-shell” (e.g., Cr, Mn, Fe, Cu, Zr, Hf).
  • a "Poor Metal” refers to metals that are considered in the art to include some metallic elements of the p-block in the periodic table which are more electronegative than transition metals, and, as defined by the Los Alamos National Laboratory, include "post-transition metals" which include "Al, Ga, In, Tl, Sn, Pb and Bi. As their name implies, they have some of the characteristics of the transition elements.
  • Poor Metals can also include metalloids, which include "B, Si, Ge, As, Sb, Te, and Po. They sometimes behave as semiconductors (B, Si, Ge) rather than as conductors.” These various metal tungstates are generally synthesizable in the micro or nano- sized particulate form.
  • radio-luminescent materials also include metal molybdates, i.e., M x (Mo0 4 ) y where the metal component (M) can be any compound selected from the "Alkaline Earth Metal", “Transition Metal” or “Poor Metal” group of elements in the periodic table, or an atomic mixture thereof.
  • Metal tungstates and metal molybdates share common tetrahedral atomic coordination geometries and thus similar electronic band structures; as a result, they exhibit similar radio-luminescent characteristics.
  • Metal molybdates are also synthesizable in micro/nanoparticulate form.
  • any non-toxic concentration of radio-luminescent particles can be used.
  • calcium tungstate particles or particle aggregates at a calcium tungstate concentration less than about 1.0 mg/cc of tumor can be used.
  • calcium tungstate particles or particle aggregates at any calcium tungstate concentration in the range between about 0.1 and 100 mg/cc of tumor can be used because the particles are non-toxic.
  • the mean diameter of the particles may be about 0.01, 0.1, 1, 10 or 100 ⁇ . In a particular embodiment, the mean diameter of the particles is ranged between about 1 and 100 ⁇ . In another embodiment, the mean diameter of the particles is ranged between 0.01 and 1 ⁇ .
  • the radio-luminescent material can be a nanoparticle.
  • the mean diameter of the nanoparticles can range between about 0.1 and about 500 nm. In one example, the mean diameter of the nanoparticles may be about 0.1, 0.5, 1, 5, 10, 50, 100, 150, 200 or 500 nm.
  • any radio-luminescent metal tungstate or molybdate particle with a mean largest dimension between 0.001 and 50 ⁇ (as determined in the unaggregated state) can be used.
  • Any radio-luminescent metal tungstate or molybdate particle aggregate with a mean largest dimension between 0.001 and 500 ⁇ can be used.
  • the radio-luminescent metal tungstate/molybdate particle or particle aggregate has a mean diameter in the range between 0.001 and 0.5 ⁇ .
  • the radio-luminescent particles of the invention emit UV light in response to high- energy photon radiation.
  • high-energy photon radiation include, but not limited to, gamma ( ⁇ ) ray and X-ray radiations.
  • the radio-luminescent particles can also be excited by other types of ionizing radiation, including electron beam radiation and alpha/beta particles produced during radioactive decay of some radioactive isotopes.
  • the radio-sensitization method based on radio-luminescent particles works well with clinically relevant radiations with MV-level X-ray/y-ray photon energies.
  • the radio-luminescent particles are used with ionizing radiation with an energy in the range of between about 1 keV and about 50 MeV.
  • the radio-luminescent particles of the invention can be conjugated or operably linked to a targeting agent ("ligand") specific to cancer cells.
  • a targeting agent ligand
  • lung cancer specific targeting ligand can be conjugated to the surfaces of the radio-luminescent particles in order to enhance the delivery of the particles to lung cancer cells.
  • the targeting agent can be a biological molecule (e.g., an antibody) having specific affinity to cancer cells.
  • cancer targeting agents include, but not limited to, folic acid, transferrin, and monoclonal antibodies against CD123, CD33, CD47, CLL-1 , etc. Any suitable conjugation or linking method, known to one of skilled in the art, can be used.
  • the radio-luminescent particles or particle aggregates of the invention can be encapsulated within a biocompatible coating material.
  • the coating material can be a polymer, for example, a biodegradable or biocompatible amphiphilic block copolymer. Any suitable polymer can be used for this purpose. Examples of polymers include, but are not limited to, polyethylene glycol (PEG), poly(D,L-lactic acid) (PLA), poly(D,L-lactic acid-ran- glycolic acid) (PLGA), poly(s-caprolactone) (PCL), poly(styrene) (PS), or a copolymeric combination of two or more of these polymer components.
  • the coating material itself can be functionalized with a cancer-targeting moiety such as folic acid.
  • a method of treating cancer in a subject comprising: providing radio-luminescent particles to a tumor site associated with said cancer in said subject; and exposing said tumor cells to high-energy ionizing radiation, wherein said particles emit UV light in response to said ionizing radiation, thereby treating said cancer in said subject.
  • the treatment may improve tumor cell killing without increasing toxicity to normal tissue.
  • the tumor to be treated can be a solid or hematological tumor.
  • Said subject can be a human or an animal (e.g., an animal patient presented to an animal hospital). Possible routes of administration of the radio-luminescent particle include, but not limited to, intratumoral, intravenous, intraarterial, and intraperitoneal.
  • the terms “treat” and “treatment” refer to therapeutic treatment, including prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) undesirable physiological changes associated with a disease or condition.
  • Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of the extent of a disease or condition, stabilization of a disease or condition (i.e. , the state in which the disease or condition does not worsen), delay or slowing down of the progression of a disease or condition, amelioration or palliation of the disease or condition, and remission (whether partial or total) of the disease or condition, whether detectable or undetectable.
  • Treatment can also mean prolonging survival as compared to expected survival if a patient were not receiving treatment.
  • Those in need of treatment include those already with the disease or condition as well as those prone to having the disease or condition or those in which the disease or condition is to be prevented.
  • the terms “treat” and “treatment” refer to inhibiting tumor growth.
  • the method of the invention can be used to treat any cancer/tumor.
  • cancer/tumor types which may be treated include, but not limited to, head and neck cancer, breast cancer, prostate cancer, lung cancer, lung cancer, gynecological cancer, cervical cancer, brain cancer, melanoma, and colorectal cancer (including HER2+ and metastatic).
  • Additional examples of cancers/tumors which may be treated include, but not limited to, bladder cancer, ovarian cancer, and gastrointestinal cancer.
  • Examples of lung cancer include, but are not limited to small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC).
  • Cancers/tumors that can be treated include primary, secondary and metastatic tumors (including those metastasized from lungs, breast, prostate, gastrointestinal tract, kidney, and larynx) as well as recurrent or refractory tumors.
  • Recurrent tumors encompass tumors that appear to be inhibited by treatment but recur up to five years, sometimes up to ten years or longer after treatment is discontinued.
  • Refractory tumors are tumors that have failed to respond or are resistant to treatment with one or more conventional therapies for the particular tumor type.
  • Refractory tumors include those that are hormone-refractory (e.g., androgen-independent prostate cancer, or hormone-refractory breast cancer such as breast cancer that is refractory to tamoxifen), those that are refractory to treatment with one or more chemotherapeutic agents, those that are refractory to radiation therapy, and those that are refractory to combinations of chemo and radiation therapies, chemo and hormone therapies, or hormone and radiation therapies.
  • hormone-refractory e.g., androgen-independent prostate cancer, or hormone-refractory breast cancer such as breast cancer that is refractory to tamoxifen
  • chemotherapeutic agents those that are refractory to radiation therapy
  • combinations of chemo and radiation therapies chemo and hormone therapies, or hormone and radiation therapies.
  • Therapy may be "first-line", i.e. , as an initial treatment in a patient who has had no prior anti-cancer treatments, either alone or in combination with other treatments, or "second-line", as a treatment in a patient who has had one prior anti-cancer treatment regimen, either alone or in combination with other treatments, or as "third-line,” “fourth-line,” etc., treatments, either alone or in combination with other treatments.
  • Therapy may also be given to a patient who has had previous treatments which have been partially successful but is intolerant to the particular treatment. Therapy may also be given as an adjuvant treatment, i.e. , to prevent reoccurrence of cancer in a patient with no currently detectable disease or after surgical removal of tumor.
  • a method for improving the effectiveness of radiation therapy for treatment of cancer in a subject comprising: providing radio- luminescent particles or particle aggregates to a tumor site associated with said cancer in said subject; and exposing said tumor cells to high-energy ionizing radiation, wherein said particles emit UV light in response to said ionizing radiation, thereby improving the effectiveness of said radiation therapy for treatment of said cancer in said subject.
  • a method for sensitizing tumor cells to radiation therapy for treatment of cancer in a subject comprising: providing radio- luminescent particles or particle aggregates to said tumor; and exposing said tumor cells to ionizing radiation, wherein said particles emit UV light in response to said ionizing radiation, thereby sensitizing said tumor cells to said radiation therapy for treatment of said cancer in said subject.
  • a method for improving tumor cell killing for treatment of cancer in a subject comprising: providing radio-luminescent particles or particle aggregates to a tumor site associated with said cancer in said subject; and exposing said tumor cells to ionizing radiation, wherein said particles emit UV light in response to said ionizing radiation, thereby improving tumor cell killing for treatment of said cancer in said subject.
  • a method for generating secondary UV radiation in deep tissue tumors comprising delivering radio-luminescent particles or particle aggregates to said tumor and illuminating said particles with deep-penetrating radiation (e.g., ⁇ rays or X-rays), thereby generating said secondary UV radiation in said tumor.
  • deep-penetrating radiation e.g., ⁇ rays or X-rays
  • Radio-Luminescent Particles for Enhancement of Radiation Cancer Therapy are Radio-Luminescent Particles for Enhancement of Radiation Cancer Therapy
  • Cancer is one of the leading causes of death worldwide. Current data suggest that radiation and concurrent chemotherapy may increase survival of patients with inoperable cancers. However, overall progress in radiation treatment of late stage cancer is still limited. Here, we demonstrate a novel approach for improving the efficiency of radiation therapy in the treatment of cancer without increasing normal tissue toxicity— namely, by simultaneous addition of radio-luminescent particles (which emit UV light under high energy photon radiation), to ⁇ ray/X-ray radiation treatments.
  • UV light with ionizing radiation such as ⁇ rays and X-rays
  • ionizing radiation such as ⁇ rays and X-rays
  • UV has never before been thought useful for radiation sensitization in clinical radio-therapy because UV light has a very limited penetration distance in tissue ( ⁇ 1 mm in human tissue).
  • Our recent work demonstrates that one can combine radiation therapy with UV treatment by utilizing radio-luminescent agents which emit UV light as a result of exposure to ionizing radiation; therefore, secondary UV radiation can be generated even in deep tissue tumors by delivering radio-luminescent particles to the tumor and illuminating them with deep-penetrating ⁇ rays or X-rays ( Figure 1).
  • CaWCU calcium tungstate
  • Concurrent UV irradiation produces sensitization effects because DNA damage by UV light may likely initiate repair sequence, and arrest progression of the cell cycle from G2 to mitosis; cancer cells are likely most susceptible to radiation damage when they are in the G2/M phase.
  • the un-optimized radio-luminescent material is already able to induce about a factor of two enhancement of the cytotoxic effect of ⁇ rays, a level of increase not easily seen with conventional radio-sensitizers.
  • the sensitization effect may become even greater when CaWCU is used in nanoparticle form; smaller nanoparticles have larger surface areas and can produce higher induced UV emissions; also, nanoparticles can easier be internalized by cancer cells.
  • CaWCU radio-luminescent nanoparticles having various sizes in the range of 20 to 200 nm can be first synthesized by the solvothermal reaction method.
  • the detailed atomic structures of these CaWCU nanoparticles can be determined by X-ray diffraction and transmission electron microscopy.
  • CaW0 4 nanoparticles can be surface functionalized by encapsulation with poly(lactic acid-co-glycolic acid)-poly(ethylene glycol) or poly(lactic acid- co-glycolic acid)-poly(ethylene glycol)-folate.
  • the size and size distribution characteristics of the uncoated, PEG-encapsulated, and folate-functionalized CaW0 4 nanoparticles can be characterized by dynamic light scattering and analytical ultracentrifugation. How size and surface characteristics affect the luminescence properties of CaW0 4 RLNP can be evaluated under X-ray excitation by spectrophotometry. The effects of size and surface chemistry on the cellular internalization and intracellular trafficking of the nanoparticles can also be investigated in human lung cancer cell lines by flow cytometry and confocal microscopy.
  • CaW0 4 nanoparticles can be prepared by the solvothermal reaction of sodium tungstate dihydrate (Na2W0 4 -2H20) with calcium salt (such as calcium chloride (CaCl2-2H20), calcium nitrate (Ca(N03)2-4H20) or calcium acetate (Ca(CH3COO)2)).
  • Surfactant such as CTAB, Triton X or PEG
  • a hydrothermal post-treatment method can be used to fine tune the sizes of final CaW0 4 materials to be studied.
  • the radio-luminescence of CaW0 4 can be determined by the crystallinity of the material.
  • the detailed structural characterizations can be performed using the X-ray diffraction (XRD) and high-resolution transmission electron microscopy (HR-TEM) techniques.
  • XRD X-ray diffraction
  • HR-TEM high-resolution transmission electron microscopy
  • the cellular internalization and intracellular trafficking of CaW0 4 nanoparticles can be significantly influenced not only by the mean size but also by the size distribution of the nanoparticles.
  • the mean particle size data can be obtained using the dynamic light scattering (DLS) technique.
  • the precise size distribution characteristics of the CaW0 4 nanoparticles can be examined using the analytical ultracentrifugation (AUC) technique.
  • AUC analytical ultracentrifugation
  • CaW0 4 nanoparticles Surface functionalization of CaW0 4 nanoparticles with PEG and folic acid.
  • the as-synthesized CaW0 4 particles can be further stabilized by encapsulation with a biodegradable block copolymer (BCP), poly(lactic acid-co-glycolic acid) -poly (ethylene glycol) (PLGA-PEG), via the solvent exchange procedure.
  • BCP biodegradable block copolymer
  • PLGA-PEG poly(lactic acid-co-glycolic acid) -poly (ethylene glycol)
  • DLS can be used to measure the mean size, and AUC to determine the size distribution of the BCP-coated CaW0 4 nanoparticles.
  • Folate surface functionalized CaW0 4 nanoparticles can also be prepared.
  • the folate receptor is overexpressed in many cancel cell types including lung cancer cells.
  • folate ligands can enhance the internalization of the nanoparticles in cancer cells.
  • a folate-functionalized PLGA-PEG BCP can be synthesized using the fluorenylmethyloxycarbonyl chloride (FMOC) protection chemistry described in our previous publication.
  • radio-luminescence/phosphorescence properties (emission wavelength, luminescence timescale, etc.) of CaW0 4 nanoparticles of various sizes (20 - 200 nm) and surface functionalities (uncoated, PEG-encapsulated, and folate-functionalized) can be characterized by spectrophotometry under the specific X-ray excitation conditions that can be used in Example 3 studies described below (i.e., 250 kVp and 6 MV X-rays).
  • the effect of PEG/folic acid surface functionalization on cellular uptake of CaWCU nanoparticles can be studied using the model lung cancer cell lines described in Example 3 and the fluorescence- activated cell sorting (FACS) technique. Intracellular trafficking properties can be analyzed in the lung cancer cells by confocal microscopy, known in the art. For these studies, a fluorescently labeled (AlexFluor647) version of the PLGA-PEG block polymer can be prepared using the FMOC protection method.
  • RLNP radio-luminescent nanoparticles
  • UV irradiation inhibits cell growth of small cell lung cancer (SCLC) and non- small cell lung cancer (NSCLC) cell lines and/or alter cell cycle distribution can be first investigated. If this inhibition is p53-dependent or involves the induction of apoptosis can be determined. One can then test whether UV light generated by CaW0 4 RLNP under X-ray radiation increases the biological effectiveness after small and large radiation fractions and determine whether any observed enhancement is dependent on the size and surface characteristics of the CaW0 4 RLNP which can determine the cellular uptake rate and intracellular location of the nanoparticles.
  • SCLC small cell lung cancer
  • NSCLC non- small cell lung cancer
  • kinetics of DNA double strand break repair and amount of residual unrepaired DNA damage can be determined by pulse field gel electrophoresis and the comet assay by established methods after large radiation fraction sizes (10, 15, 20, and 40 Gy) and one can test whether any of the above endpoints are dependent on the constitutive level of NF- ⁇ in the cells.
  • NF- ⁇ can be determined by gel shift assays according to methods known in the art.
  • CaW0 4 RLNP inhibits cell growth of SCLC and NSCLC cell lines or alters cell cycle distribution. Separately, one can also investigate whether UV irradiation alone causes growth inhibition of the SCLC and NSCLC cell lines and/or alter cell cycle distribution. Whether this inhibition is p53- dependent or involves the induction of apoptosis can be determined. One can then determine whether CaW0 4 LRNP increases the biological effectiveness after small and large radiation fractions by the published assays listed above.
  • any observed enhancement is dependent on the size and surface characteristics of the nanoparticles which can determine their cellular uptake rates and intracellular distributions. Whether the radio- sensitization effect of CaW0 4 RLNP is related to the inhibition of the level of constitutive and/or radiation-induced NF- ⁇ activity in the various lung cancer cell lines can be investigated and whether double strand break DNA repair is altered after small or large fraction X-ray irradiation in the presence (and absence) of CaW0 4 RLNP can be determined.
  • Lung cancer is the leading cancer death in the United States and worldwide.
  • radiation and concurrent chemotherapy such as cisplatin and etoposide may increase median survival in inoperable Stage III NSCLC with good performance status.
  • chemo-radiation also appears to have some benefit; however, overall progress is still limited.
  • a novel concept that the addition of concurrent, tumor-targeted radio-luminescent nanoparticles (which emit UV light under high energy photon radiation), to fractionated radio-therapy approaches may improve tumor cell killing without increasing normal tissue toxicity.
  • the preclinical studies can determine whether the biological efficacy of fractionated radiation treatments can be enhanced by concurrent treatment with this new type of radio-sensitizer, CaW0 4 radio-luminescent nanoparticles (RLNP), and can also determine the detailed biological mechanism of the UV-induced radio- sensitization effect, i.e., whether this enhancement is due to alteration of the mode of cell death or DNA repair, and whether NSCLC or SCLC respond in a similar manner.
  • RLNP radio-luminescent nanoparticles
  • Radio-Luminescent Particles As a means to achieve radio-sensitization and study the ability to affect head and neck cancer cells (in vitro) and xenografts (in vivo).
  • This Radio Luminescence Therapy (RLT) technology enables cancer patients to achieve the benefits of radiation treatment with reduced negative effects.
  • Radiation therapy is one of the three pillars of cancer therapy alongside chemotherapy and surgery. About two-thirds of all cancer patients receive radiation therapy during their illness, and in the US annually nearly a million patients are treated with radiation therapy.
  • radiation therapy generally carries significant side effects in patients, both acute and chronic, because of the damage high-energy ionizing radiation (such as X-ray or ⁇ ray radiation) causes to normal cells and tissues. Due to the side effects, patients receive small amounts of radiation over weeks of period which increases cost of treatment not only for the patients but also for hospitals and insurance companies; in the US, hospitals annually spend $5.8 billion in radiation equipment, and insurance companies annually spend $15 billion in
  • radiotherapy is also an economic burden to the entire health care system.
  • Radio-sensitizers that make cancer cells easier to kill with radiation therapy.
  • Many anticancer drugs have radio-sensitization effects.
  • Some drugs such as doxorubicin, 5- fluorouracil, cisplatin
  • sensitize cancer cells to radiation by intercalation with DNA and others (e.g., paclitaxel, etanidazole) produce sensitization effects by arresting the cell cycle, usually in the G2/M phase.
  • chemo radio therapy has now become standard care for some cancers.
  • Nanoparticles offer improvements, because, due to their larger sizes, nanoparticles are easier to maintain at the tumor sites for longer periods of time.
  • metal/metal oxide nanoparticles these "high-Z" materials (particularly, those derived from gold, silver, iron, gadolinium, hafnium) produce strong secondary electron radiation due to the photoelectric, Compton and Auger effects, and thus cause localized augmentation of radiation damage.
  • the sensitizing effects of these currently available compounds are not satisfactory; none of these previous methods is able to increase the radiation's potency by more than a factor of about 1.5.
  • these photo-electric nanoparticles are limited because of the small mean free paths of electrons.
  • Nanoparticles that produce photons (instead of electrons) with X-ray should be better materials for radio- sensitization, because photons have two to three orders of magnitude larger mean free paths than electrons.
  • current radio-sensitization methods based on photo-electric nanoparticles work best with X-rays of the order of 100 kVp in energy, but not as well with more clinically relevant radiations with MV-level X-ray/y-ray photon energies because of the significantly reduced absorption cross-sections at the higher energies.
  • the most studied material in this regard i.e., nanoparticulate gold, is known to cause genotoxic and mutagenic effects in exposed tissues in vivo.
  • Radio Luminescence Therapy (RLT)
  • RLT Radio Luminescence Therapy
  • This method utilizes a new type of radio- sensitizers, namely, "Radio-Luminescent Particles (RLPs)".
  • RLPs emit UV light, instead of secondary electrons, as a result of exposure to ionizing radiation (such as X-ray or ⁇ ray).
  • ionizing radiation such as X-ray or ⁇ ray.
  • X-ray and UV light significantly enhances the cancer cell-killing effectiveness of X-rays.
  • This RLT works even better with clinically relevant MV- level X-rays.
  • This RLT technology can be applied to the animal radio- sensitizer market (for companion animals with malignant cancers.
  • UV light itself has genotoxic effects on cancer cells; it causes damage to DNA in cancer cells. DNA damage by UV light initiates repair sequence, and arrests progression of the cell cycle from G2 to mitosis. Therefore, it is beneficial to combine radiation therapy with UV treatment, because cancer cells are most susceptible to radiation damage when they are in the process of separating the replicated chromosomes during cell division (i.e., in the G2/M phase). This is the underlying hypothesis which the herein disclosed RLT technology is based upon. Synergistic interactions of UV light with X-rays in killing cells have been known for more than 50 years.
  • UV has never before been thought useful for X-ray/ ⁇ ray sensitization in clinical radiation therapy because UV light has a very limited penetration distance in tissue ( ⁇ 1 mm).
  • work by our laboratory demonstrates that it is possible to combine radiation therapy with UV treatment by utilizing radio-luminescent agents which emit UV light under ionizing radiation (Figure 1).
  • Secondary UV radiation can be generated even in deep tissue tumors by delivering radio-luminescent particles to the tumor and illuminating them with deep-penetrating ⁇ rays or X-rays. This is a new paradigm in cancer radiation therapy.
  • the herein disclosed technology utilizes a naturally- abundant radio-luminescent mineral, calcium tungstate (CaW0 4 ) in its micro or nanoparticulate form for generating secondary UV light by ⁇ ray/X-ray photons.
  • This CaW0 4 material exhibits a luminescence emission with a maximum at 420 nm wavelength under high energy ionizing radiation such as ⁇ rays, X-rays or short wavelength UV light at room temperature ( Figures 5A - 5G).
  • Our RLP formulations are designed to be non-toxic to the human body.
  • the RLP core is composed of CaW0 4 , which is chemically stable and non- toxic, and this CaW0 4 core is further coated with an FDA-approved polymer (poly(ethylene glycol-block-D,L-lactic acid) (PEG-PLA)) to make it inert to proteins; encapsulated CaW0 4 (CWO) has previously been shown to possess no detectable cytotoxicity against HeLa cells. Unlike gold or silica, CaW0 4 does not have any reactive sites, and the PEG surface functionalization can be achieved only by the method developed by our laboratory. Both nanoparticle fabrication and encapsulation processes are scalable for production of large quantities. These RLPs are formulated in solution form, and can be injected into the tumor without loss.
  • FDA-approved polymer poly(ethylene glycol-block-D,L-lactic acid) (PEG-PLA)
  • PEG-PLA poly(ethylene glycol-block-D,L-lactic acid)
  • CWO encapsulated CaW0 4
  • CaW0 4 does not
  • Particle size and surface functionality are two important parameters that need to be optimized in order to maximize the intratumoral retention and distribution, the cellular internalization, and thus the overall radio-sensitization efficiency, and also the eventual pharmacokinetic fate of the material can be determined by these parameters.
  • RLPs can be used in human cancer patients. For regulatory reasons, we will pursue the animal market first.
  • the product will be RLPs in a solution that can be injected directly to tumor prior to radiation treatment; there is no need to change the existing radiation-treatment platform.
  • CWO NPs block copolymer(BCP)-encapsulated CaW0 4 (CWO) nanoparticles
  • the CWO NPs were synthesized by a micro-emulsion method. First, 20 ml of cyclohexane was mixed with 2 ml of hexanol. CTAB (2 mmol) (> 99%, Sigma) was added to this solvent mixture, and then the solution was heated to 70 °C or until the solution became transparent (Solution 1). Meanwhile, 0.4 mmol of Na2W0 4 (99%, Acros Organics) was dissolved in 0.6 ml of Milli-Q water (Solution 2).
  • the PEG-PLA diblock copolymer was synthesized by l,8-diazabicyclo[5.4.0] undec- 7-ene(DBU, 98%, Aldrich)-catalyzed ring-opening polymerization of lactide (LA, a racemic mixture). 0.45 g of PEG-ME was dissolved in DCM (22 ml) dried with molecular sieves. After a day LA (0.35 g) was added into the PEG-ME solution. The polymerization was initiated by adding 2 ml of a DBU solution (3.35 mmol of DBU dissolved in 30 ml of DCM) to the LA/PEG-ME mixture at room temperature.
  • the polymerization reaction was run for 1 hour at room temperature. Afterward the reaction was terminated by adding 10 mg of benzoic acid (> 99.5%, Sigma- Aldrich). The polymerization mixture was added drop-wise to 1000 ml petroleum ether for precipitation. After the PEG-PLA product settled to the bottom, the supernatant was decanted. The polymer was dried in a vacuum oven.
  • PEG-PLA-encapsulated CWO samples were prepared as follows. 1.0 mg of CWO (purified by centrifugation) was dispersed in 1.0 g of DMF. 100.0 mg of PEG-PLA was added to 2.9 g of the above nanoparticle suspension. This mixture was stirred using a high speed overhead mechanical stirrer (at 15000 rpm) with simultaneous sonication. 2.1 ml of Milli-Q water was added to the DMF solution. The resulting mixture was emulsified with a mechanical stirrer and then ultrasonicated in a sonication bath for 30 minutes.
  • This emulsion was placed in a dialysis bag (molecular weight cutoff 50 kDa) and dialyzed for 3 days against a total of 1.0 liter of Milli-Q water (regularly replaced with fresh Milli-Q water) to remove DMF.
  • HN31 cells were cultured in DMEM (Life Technologies, Carlsbad, CA, USA) supplemented with 10% FBS, 100 Units/ml penicillin and 100 ⁇ g/ml streptomycin in humidified atmosphere with 5% CO2 37°C.
  • FBS fetal bovine serum
  • penicillin 100 Units/ml penicillin
  • streptomycin 100 ⁇ g/ml streptomycin in humidified atmosphere with 5% CO2 37°C.
  • CWO CaWC
  • the cells were treated with uncoated CWO microparticles (MPs) or PEGylated CWO nanoparticles (NPs) for 24 h at the various CWO concentrations indicated in Figure 6.
  • MPs uncoated CWO microparticles
  • NPs PEGylated CWO nanoparticles
  • tumors were grown over a 21-day period to approximately 250 mm 3 , total 0.30 mg of uncoated CWO MPs or PEG-PLA-encapsulated CWO NPs were infused directly into the tumor (total 1.2 mg of CWO per cc of tumor); the procedure involved two injections of 120 ⁇ CWO solution in PBS (CWO concentration: 1.23 mg/ml) over a two-day period.
  • tumor volume was measured for 54 days (using the formula (TI/6)XLXWXH to give volume in ml).
  • CaWCU (CWO) RLPs of various sizes (2 - 100 nm diameters) have been successfully prepared by the reaction of sodium tungstate dihydrate with calcium salt ( Figures 5A - 5G).
  • Surfactant was used to control the particle nucleation and growth kinetics.
  • a post-reaction thermal treatment method was used to fine-tune the final size.
  • the radio-luminescence of CaWCU is due to the unique crystal structure and resulting electronic band gap of the material.
  • Detailed structural characteristics (grain size, shape, and crystallinity) of the synthesized CaWCU nano crystals were characterized using the X-ray diffraction and high-resolution TEM techniques ( Figures 5B and 5C).
  • the as-synthesized CaWCU NPs were coated with different surfactant materials (e.g., citric acid, cetyl trimethylammonium bromide, a mixture of oleic acid and oleylamine).
  • surfactant materials e.g., citric acid, cetyl trimethylammonium bromide, a mixture of oleic acid and oleylamine.
  • the original surfactant coating could be replaced with a new enclosure formed by block copolymer (BCP) materials using the method developed by our laboratory.
  • BCP block copolymer
  • Two types of BCPs have been tested: PEG-PLA, and poly(ethylene glycol- block-n-butyl acrylate) (PEG-PnBA).
  • the values of the Sensitization Enhancement Ratio or SER (defined as the ratio of the radiation dose at 10% clonogenic survival in the absence of CWO relative to the radiation dose at 10% survival in the presence CWO) were estimated to be 1.15 for MP + ⁇ (0.5 mg/ml CWO) and 1.13 for NP + ⁇ (0.125 mg/ml CWO).
  • the / ⁇ -values relative to control (“ ⁇ ") were estimated to be 0.13 at 3 Gy, 0.04 at 6 Gy, and 0.04 at 9 Gy.
  • the p-values relative to ⁇ were 0.03 at 3 Gy, 0.11 at 6 Gy, and 0.06 at 9 Gy.
  • the radio-sensitization effect appears to be not related to the cellular uptake and intratumoral location of the CWO RLPs because the CWO MPs should not have been internalized by the tumor cells due to their large size. It should be noted that in both these initial in vitro clonogenic and in vivo tumor growth assays ( Figures 8 and 9, respectively) we used intratumoral CWO RLP concentrations that were at least five-fold lower than the target clinical CWO dose level of intratumoral 5 - 10 mg per cc of tumor; it is remarkable that despite the small amounts of CWO used significant radio-sensitization effects of CWO RLPs were clearly visible.
  • the above clinical dose value (i.e., 5 - 10 mg CWO/cc of tumor) was estimated using the following information.
  • CaW0 4 has a radio-luminescence energy transfer efficiency of about 5% for X-ray/ ⁇ ray radiation.
  • each cell should receive 6.3 x 10 12 nanoparticles.
  • the cell density within a solid tumor is known to be about 1.7 x 10 5 cells per cc of tumor
  • the above information now translates into the likely minimum amount of CWO NPs to be administered by direct intratumoral infusion into the tumor, that is, about 3.3 mg CWO per cc of tumor. Therefore, in the clinical embodiment of the RLT technology, it is desirable that about 5-10 mg of CWO NPs per cc of tumor be infused into the tumor prior to delivery of the first fraction of radiation therapy. Preclinical testing of this protocol is also currently underway.
  • the folate receptor is overexpressed not only in many cancer cell types including head and neck cancer cells but also in immunosuppressive tumor-associated macrophages; folate ligands will enhance the internalization of the nanoparticles in both of these cell types.
  • intratumoral nanoparticle injections and localized radiation therapy our proposed therapy will have three arms of potential effectiveness, namely, radiotherapy, UV therapy, and immunotherapy. Therefore, the size and surface functionality need to be optimized for maximum radio-sensitization efficiency.
  • Radio-Luminescent Particles namely, "Radio-Luminescent Particles (RLPs)
  • RLPs Radio-Luminescent Particles

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Abstract

L'invention se rapporte de manière générale à un procédé de radiosensibilisation qui permet d'améliorer l'efficacité de la radiothérapie pour le traitement du cancer. Spécifiquement, l'invention concerne l'utilisation de particules radioluminescentes pour sensibiliser les cellules tumorales à la radiothérapie.
PCT/US2016/012616 2015-01-08 2016-01-08 Particules radioluminescentes pour l'amélioration de la radiothérapie du cancer Ceased WO2016112268A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020154110A1 (fr) * 2019-01-22 2020-07-30 Purdue Research Foundation Particules radio-luminescentes revêtues de bilirubine

Families Citing this family (1)

* Cited by examiner, † Cited by third party
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080003183A1 (en) * 2004-09-28 2008-01-03 The Regents Of The University Of California Nanoparticle radiosensitizers
US20110021970A1 (en) * 2007-11-06 2011-01-27 Duke University Non-invasive energy upconversion methods and systems for in-situ photobiomodulation
US20130116616A1 (en) * 2010-07-17 2013-05-09 Merck Patent Gmbh Enhancement of penetration and action
US20140272030A1 (en) * 2007-04-08 2014-09-18 Immunolight, Llc. Interior energy-activation of photo-reactive species inside a medium or body

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5407659A (en) * 1991-10-22 1995-04-18 Mallinckrodt Medical, Inc. Treated calcium/oxyanion-containing particles for medical diagnostic imaging
US20040058457A1 (en) * 2002-08-29 2004-03-25 Xueying Huang Functionalized nanoparticles
FR2869803B1 (fr) * 2004-05-10 2006-07-28 Nanobiotix Sarl Particules activables, preparation et utilisations
WO2007149951A2 (fr) * 2006-06-20 2007-12-27 Sunstone Inc. Systèmes et procédés d'utilisation de composés luminescents dans le traitement de maladies et l'imagerie médicale
US9072789B2 (en) * 2007-07-20 2015-07-07 The Trustees Of Princeton University Nano-particle surface modification
EP2130553A1 (fr) * 2008-06-05 2009-12-09 Nanobiotix Nanoparticules inorganiques de haute densité pour détruire des cellules in-vivo
WO2013131064A1 (fr) * 2012-03-01 2013-09-06 Ferro Corporation Composés d'absorption laser
US9700632B2 (en) * 2012-09-13 2017-07-11 Centre For Bioseperation Technology-Vit Dendrimers, conjugates and methods thereof
US11337665B2 (en) * 2013-03-15 2022-05-24 The Trustees Of The University Of Pennsylvania Radiographic contrast agents for temporal subtraction and dual-energy x-ray imaging
US20160220500A1 (en) * 2014-11-14 2016-08-04 Kent State University Targeting Intracellular Copper Ions for Inhibiting Angiogenesis Using Nanoparticles of Ternary Inorganic Metal Sulfide M1M2S4 (M1, independently, is Mg, Ca, Mn, Fe, or Zn; M2 = Mo or W) Compounds to Treat Metastatic Cancer

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080003183A1 (en) * 2004-09-28 2008-01-03 The Regents Of The University Of California Nanoparticle radiosensitizers
US20140272030A1 (en) * 2007-04-08 2014-09-18 Immunolight, Llc. Interior energy-activation of photo-reactive species inside a medium or body
US20110021970A1 (en) * 2007-11-06 2011-01-27 Duke University Non-invasive energy upconversion methods and systems for in-situ photobiomodulation
US20130116616A1 (en) * 2010-07-17 2013-05-09 Merck Patent Gmbh Enhancement of penetration and action

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020154110A1 (fr) * 2019-01-22 2020-07-30 Purdue Research Foundation Particules radio-luminescentes revêtues de bilirubine
US20220127528A1 (en) * 2019-01-22 2022-04-28 Purdue Research Foundation Bilirubin-coated radio-luminescent particles

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WO2016112314A1 (fr) 2016-07-14
US20180008733A1 (en) 2018-01-11

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